Simulation of thermal ablation by high-intensity focused ultrasound with temperature-dependent properties.

An integrated computational framework was developed in this study for modeling high-intensity focused ultrasound (HIFU) thermal ablation. The temperature field was obtained by solving the bioheat transfer equation (BHTE) through the finite element method; while, the thermal lesion was considered as a denatured material experiencing phase transformation and modeled with the latent heat. An equivalent attenuation coefficient, which considers the temperature-dependent properties of the target material and the ultrasound diffraction due to bubbles, was proposed in the nonlinear thermal transient analysis. Finally, a modified thermal dose formulation was proposed to predict the lesion size, shape and location. In-vitro thermal ablation experiments on transparent tissue phantoms at different energy levels were carried out to validate this computational framework. The temperature histories and lesion areas from the proposed model show good correlation with those from the in-vitro experiments.

Induced depressive behavior impairs learning and memory in rats.

While it is generally accepted that cognitive processes such as learning and memory are affected by emotion, the impact of depression on learning and memory has rarely been directly studied in experimental animals. Effects of induced depressive behavior on learning and memory were determined in rats, using an open space swim test, a novel animal model of depressive behavior that is developed recently in our laboratory. The model indexes searching activity of the animals, with the induced depressive immobility behavior showing specific sensitivity to three major prototypic classes of antidepressants and a selective serotonin reuptake inhibitor. The induced depressive behavior in rats showed a delayed response to chronic antidepressant treatment and had a lasting effect on the ability of rats to learn and recall the learned experience. It impaired the subsequent ability of rats to learn and recall both a spatial water maze task and a multi-trial passive avoidance task. These impairments were all sensitive to antidepressant therapeutics, but not to buspirone, an anxiolytic. By way of contrast, the ability of the rats to sense and move to a visible platform and to escape from an unconditioned shock stimulus was neither impaired by inducing the depressive behavior nor altered by the drug treatment, suggesting that non-specific changes in sensorimotor ability were not involved. These impairments of learning and memory indicate that the depressive behavior-induced deficits show generalizability and are not context-limited. This animal model of depressive behavior shows promising potential as a screen for novel antidepressive therapeutics and as a disease model for revealing network/cellular/molecular mechanisms in the pathophysiology of depression and depression-induced cognitive deficits.

Regulation of NMDA receptors by cyclin-dependent kinase-5.

Members of the N-methyl-d-aspartate (NMDA) class of glutamate receptors (NMDARs) are critical for development, synaptic transmission, learning and memory; they are targets of pathological disorders in the central nervous system. NMDARs are phosphorylated by both serine/threonine and tyrosine kinases. Here, we demonstrate that cyclin dependent kinase-5 (Cdk5) associates with and phosphorylates NR2A subunits at Ser-1232 in vitro and in intact cells. Moreover, we show that roscovitine, a selective Cdk5 inhibitor, blocks both long-term potentiation induction and NMDA-evoked currents in rat CA1 hippocampal neurons. These results suggest that Cdk5 plays a key role in synaptic transmission and plasticity through its up-regulation of NMDARs.

Pharmacological enhancement of synaptic efficacy, spatial learning, and memory through carbonic anhydrase activation in rats.

CA1 pyramidal cells were recorded in rat hippocampal slices. In the presence of carbonic anhydrase activators, comicrostimulation of cholinergic inputs from stratum oriens and gamma-aminobutyric acid (GABA)ergic inputs from stratum pyramidale at low intensities switched the hyperpolarizing GABA-mediated inhibitory postsynaptic potentials to depolarizing responses. In the absence of the activators, however, the same stimuli were insufficient to trigger the synaptic switch. This synaptic switch changed the function of the GABAergic synapses from excitation filter to amplifier and was prevented by carbonic anhydrase inhibitors, indicating a dependence on HCO. Intralateral ventricular administration of these same carbonic anhydrase activators caused the rats to exhibit superior learning of the Morris water maze task, suggesting that the GABAergic synaptic switch is critical for gating the synaptic plasticity that underlies spatial memory formation. Increased carbonic anhydrase activity might, therefore, also enhance perception, processing, and storing of temporally associated relevant signals and represents an important therapeutic target in learning and memory pharmacology.

Theta rhythm of hippocampal CA1 neuron activity: gating by GABAergic synaptic depolarization.

Information processing and memory consolidation during exploratory behavior require synchronized activity known as hippocampal theta (theta) rhythm. While it is well established that the theta activity depends on cholinergic inputs from the medial septum/vertical limb of the diagonal band nucleus (MS/DBv) and theta discharges of GABAergic interneurons, and can be induced with cholinergic receptor agonists, it is not clear how the increased excitation of pyramidal cells could occur with increased discharges of GABAergic interneurons during theta waves. Here, we show that the characteristic theta activity in adult rat hippocampal CA1 pyramidal cells is associated with GABAergic postsynaptic depolarization and a shift of the reversal potential from Cl(-) toward HCO(3)(-) (whose ionic gradient is regulated by carbonic anhydrase). The theta activity was abolished by GABA(A) receptor antagonists and carbonic anhydrase inhibitors, but largely unaffected by blocking glutamate receptors. Carbonic anhydrase inhibition also impaired spatial learning in a water maze without affecting other sensory/locomotor behaviors. Thus HCO(3)(-)-mediated signaling, as regulated by carbonic anhydrase, through reversed polarity of GABAergic postsynaptic responses is implicated in both theta and memory consolidation in rat spatial maze learning. We suggest that this mechanism may be important for the phase forward shift of the place cell discharges for each theta cycle during the animal's traversal of the place field for that cell.

Functional switching of GABAergic synapses by ryanodine receptor activation.

The role of the ryanodine receptor (RyR) in modifiability of synapses made by the basket interneurons onto the hippocampal CA1 pyramidal cells was examined in rats. Associating single-cell RyR activation with postsynaptic depolarization increased intracellular free Ca(2+) concentrations and reversed the basket interneuron-CA1 inhibitory postsynaptic potential into an excitatory postsynaptic potential. This synaptic transformation was accompanied by a shift of the reversal potential from that of chloride toward that of bicarbonate. This inhibitory postsynaptic potential-excitatory postsynaptic potential transformation was prevented by blocking RyR or carbonic anhydrase. Associated postsynaptic depolarization and RyR activation, therefore, changes GABAergic synapses from excitation filters to amplifier and, thereby, shapes information flow through the hippocampal network.

Calexcitin transformation of GABAergic synapses: from excitation filter to amplifier.

Encoding an experience into a lasting memory is thought to involve an altered operation of relevant synapses and a variety of other subcellular processes, including changed activity of specific proteins. Here, we report direct evidence that co-applying (associating) membrane depolarization of rat hippocampal CA1 pyramidal cells with intracellular microinjections of calexcitin (CE), a memory-related signaling protein, induces a long-term transformation of inhibitory postsynaptic potentials from basket interneurons (BAS) into excitatory postsynaptic potentials. This synaptic transformation changes the function of the synaptic inputs from excitation filter to amplifier, is accompanied by a shift of the reversal potential of BAS-CA1 postsynaptic potentials, and is blocked by inhibiting carbonic anhydrase or antagonizing ryanodine receptors. Effects in the opposite direction are produced when anti-CE antibody is introduced into the cells, whereas heat-inactivated CE and antibodies are ineffective. These data suggest that CE is actively involved in shaping BAS-CA1 synaptic plasticity and controlling information processing through the hippocampal networks.

Hypoxia, ischemic stroke, and memory deficits: prospects for therapy.

Oxygenation of cells and tissues in mammals according to metabolic needs is always a life-or-death necessity. Over the last decade, life scientists have gained an impressive understanding of cellular and molecular mechanisms in hypoxic-ischemic injury and memory deficits, survival, and death. Functional recovery from hypoxia-ischemia depends on how the individual responds to the insult, and whether appropriate responses are initiated and maintained. Unraveling the biochemical and molecular mechanisms and responses of hypoxia-ischemia is likely to dramatically shape and improve medical practice in the treatments of hypoxia-ischemia and memory deficits.

Memory and long-term potentiation (LTP) dissociated: normal spatial memory despite CA1 LTP elimination with Kv1.4 antisense.

Long-term potentiation (LTP) in the hippocampal slice preparation has been proposed as an in vitro model for long-term memory. However, correlation of LTP with memory in living animals has been difficult to demonstrate. Furthermore, in the last few years evidence has accumulated that dissociate the two. Because potassium channels might determine the weight of synapses in networks, we studied the role of Kv1.4, a presynaptic A-type voltage-dependent K+ channel, in both memory and LTP. Reverse transcription-PCR and Western blot analysis with specific antibodies showed that antisense oligodeoxyribonucleotide to Kv1.4 microinjected intraventricularly into rat brains obstructed hippocampal Kv1.4 mRNA, "knocking down" the protein in the hippocampus. This antisense knockdown had no effect on rat spatial maze learning, memory, or exploratory behavior, but eliminated both early- and late-phase LTP and reduced paired-pulse facilitation (a presynaptic effect) in CA1 pyramidal neurons without affecting dentate gyrus LTP. This presynaptic Kv1.4 knockdown together with previous postsynaptic Kv1.1 knockdown demonstrates that CA1 LTP is neither necessary nor sufficient for rat spatial memory.

Vascular effects of proteinase-activated receptor 2 agonist peptide.

Proteinase-activated receptor 2 (PAR-2) is a G protein-coupled receptor related to the thrombin receptor. PAR-2 can be activated by trypsin and by synthetic peptides corresponding to the new amino terminus generated by activating proteolytic cleavage. We show in this report that intravenous injection of PAR-2 agonist peptides has dramatic effects on arterial blood pressure in anesthetized rats. The peptide SLIGRLETQPPI, at 150 nmol/kg, transiently decreased the mean arterial pressure from 104 to 60 mm Hg. The hypotensive response was dose-dependent, and was not secondary to effects on central vasoregulatory systems, heart rate, or the kidneys. A nitric oxide synthase inhibitor attenuated the hypotensive response induced by the PAR-2 agonist peptide. Further experiments in vitro, on preparations of rat femoral artery and vein, showed that PAR-2 agonist peptide elicited a dose-dependent relaxation of both types of vessel. Removal of the endothelium abolished the agonist peptide-induced relaxation. Our results demonstrate that activation of PAR-2 can modulate vascular tone, and that this response was an effect mediated at least partly by nitric oxide. The effect on blood vessels further suggests that the physiological activator of this proteolytically activated receptor is an enzyme present and active in the blood, possibly after a vascular injury.

Ethanol inhibits chemoreflex excitation of reticulospinal vasomotor neurons.

In anesthetized and ventilated rats, activation of carotid chemoreceptors with intracarotid administration of 100 nmol sodium cyanide rapidly excited the spinal cord-projecting vasomotor neurons in the rostroventrolateral reticular nucleus (RVL) of the medulla oblongata and sympathetic nerves and increased arterial pressure. The chemoreflex sympathoexcitatory pressor responses were attenuated by an acute systemic administration of ethanol at 0.45 g/kg, but not at 45 mg/kg. The ethanol effects were observed at the level of RVL-spinal vasomotor neurons, in attenuating the neuronal responses to the chemoreflex excitation and direct iontophoresis of N-methyl-D-aspartic acid (NMDA) but without altering responses of the carotid sinus nerves to intracarotid cyanide. The effect of ethanol on the RVL neurons was further defined as blocking NMDA-evoked inward current in the corresponding spontaneously active RVL neurons in vitro. The results indicate that acute ethanol intoxication markedly influences NMDA receptor activation and arterial chemoreflexes. The relevance of the type of action to clinical hypertension in chronic and heavy drinkers is discussed.

Intracisternally applied angiotensin II does not excite reticulospinal vasomotor neurons in anesthetized rats.

We examined whether vasomotor neurons in the rostroventrolateral reticular nucleus of the medulla oblongata might be responsible for an acute increase in arterial pressure, elicited by application of angiotensin II in the central nervous system, as suggested by others. In urethane-pentobarbital-anesthetized and ventilated rats, intracisternal administration of angiotensin II (1-30 nmol, infused over a period of 30 s) produced a dose-dependent pressor response, which was abolished by intracisternal application of [Sar1, Thr8]angiotensin II (100 nmol), an angiotensin II receptor antagonist. The pressor response, however, was neither preceded by nor associated with increased discharges of vasomotor neurons with slow- and fast-conduction axons in the rostroventrolateral reticular nucleus and of lumbar sympathetic chain and renal sympathetic nerves. Intravenous injections of [beta-mercapto-beta, beta-cyclopentamethylenepropinyl1,-O-Et-Tyr2, Val4, Arg8]vasopressin, a vasopressin receptor antagonist, largely abolished the central angiotensin II-induced pressor response, while a blockade of ganglionic transmission with hexamethonium and disruption of descending sympathoexcitatory output were ineffective. We conclude that central administration of angiotensin II, under the experimental conditions and at the doses, evokes an acute pressor response largely through the release of vasopressin, not by exciting vasomotor and sympathetic neurons.

Excitatory amino acid-mediated chemoreflex excitation of respiratory neurones in rostral ventrolateral medulla in rats.

1. In anaesthetized rats, extracellular and intracellular recordings were made from 119 respiratory neurones in the rostroventrolateral reticular nucleus (RVL) of the medulla oblongata. 2. Two types of active respiratory neurones were detected in RVL: expiratory (E) and pre-inspiratory (Pre-I), based on the relationship between their discharge and that of the phrenic nerve. Some Pre-I but none of the E neurones could be antidromically excited from the C(3)-C(4) level of the spinal cord. 3. E and Pre-I neurones of RVL were excited by stimulation of the arterial chemoreceptors by a close arterial injection of sodium cyanide. The reflex excitation of RVL E neurones was preceded by increased phrenic nerve activity, while the excitation of RVL Pre-I neurones preceded the increases in phrenic nerve activity. 4. The chemoreflex excitation of the two types of RVL respiratory neurones as well as their resting discharge was abolished or significantly depressed by microionophoresis of kynurenate, a wide-spectrum antagonist of excitatory amino acid receptors, while xanthurenate, an inactive analogue of kynurenate, was without effect. 5. In ventilated rats, bilateral microinjection into RVL of kynurenate, but not xanthurenate, abolished resting activity and chemoreflex excitation of phrenic nerve activity, whilst in spontaneously breathing rats, kynurenate microinjection into RVL produced apnea and silenced phrenic nerves. 6. We conclude: (a) chemoreflex excitation of the phrenic nerves is mediated by stimulating Pre-I neurones of RVL by excitatory amino acidergic inputs and (b) RVL Pre-I neurones may directly and/or indirectly excite spinal phrenic motor neurones and hence are involved in inspiratory rhythmogenesis.

Medullary vasomotor activity and hypoxic sympathoexcitation in pentobarbital-anesthetized rats.

In pentobarbital sodium-anesthetized, paralyzed, and ventilated rats, systemic hypoxia, produced by intratracheal N2 administration for 20 s, rapidly increased activities of reticulospinal vasomotor neurons in the rostroventolateral reticular nucleus (RVL) of the medulla oblongata (by 23.9 +/- 4.7 spikes/s) and sympathetic nerves (by 30.9 +/- 4.7 microV) and arterial pressure (by 35.6 +/- 6.4 mmHg). The sympathoexcitatory and pressor responses were abolished by bilateral microinjections of muscimol, a gamma-aminobutyric acid (GABA)A-receptor agonist, (250 pmol per 50 nl/site) into the RVL. Chemical inhibition of RVL also reduced arterial pressure to 48.1 +/- 3.7 mmHg and eliminated sympathetic nerve activity. Intravenous infusion of L-phenylephrine and intrathecal administration of kainic acid restored arterial pressure to control level but not the rapid sympathoexcitatory responses to acute hypoxia. We conclude that, in pentobarbital-anesthetized rats, the sympathetic vasomotor tone and pressor responses to acute hypoxia depend on activity and excitation of RVL-spinal vasomotor neurons. The neural mechanisms responsible for the sympathetic tone and rapid pressor responses to hypoxia in these animals qualitatively differ neither from those anesthetized with urethan nor from the decerebrate unanesthetized animals.

Urethane directly inhibits chemoreflex excitation of medullary vasomotor neurons in rats.

In anesthetized rats, stimulation of the arterial chemoreceptors excites reticulospinal vasomotor neurons of the nucleus rostroventrolateral reticularis of the medulla oblongata, via activating the neuronal NMDA receptors. Excitation of these neurons is responsible for reflex increases in sympathetic neuronal activity and arterial pressure. Additional doses (0.12-0.24 g/kg i.v.) of urethane dose-dependently reduced the elevations in the discharge rate of reticulospinal vasomotor and sympathetic neurons and of arterial pressure, elicited by stimulating the carotid chemoreceptors with intra-carotid injections of sodium cyanide (100 nmol/10 microl), without affecting discharges of the carotid chemoafferents. Microiontophoresis of urethane onto the reticulospinal vasomotor neurons reversibly inhibited the excitation elicited by stimulation of the chemoreceptors and by iontophoretically applied L-glutamate. The results indicate that the suppression of chemoreflexes by excessive amount of urethane is central and one of its blocking excitatory amino acid transmission onto these vasomotor neurons.

Central neural organization and control of sympathetic nervous system in mammals.

The past decade has witnessed rapid progress in defining neural circuits and mechanisms in the brain, responsible for regulation of the sympathetic nerve activity and cardiovascular functions. Several groups of cardiovascular neurons in the brainstem form the fundamental neural circuits, through which reflexly and centrally initiated sympathetic responses are processed. Their interplay determines the levels of sympathetic nerve activity and vascular tone. Substantial evidence indicates that a small population of reticulospinal vasomotor neurons in the rostroventrolateral reticular nucleus of the medulla oblongata play critical and integrative roles by: 1) providing, largely by their intrinsic pacemaker activity, tonic sympathoexcitation, thus maintaining normal blood pressure and organ blood flows, 2) mediating a variety of circulatory reflexes and centrally initiated sympathetic responses thereby helping to match organ blood flow to metabolic demands, and 3) acting as intrinsic oxygen detectors which orchestrate appropriate autonomic response programs to protect the integrity of brain in response to acute hypoxia-ischemia. Elaboration of the neural mechanisms and cellular and molecular properties of these vasomotor neurons related to dynamic regulation of the cardiovascular system in normal and disease states will be of relevance to a full appreciation of their role in adaptation of the organism to its internal and external environments and to the development of strategies to fight against neurogenic cardiovascular diseases and to restore normal functions.

Decerebration does not alter hypoxic sympathoexcitatory responses in rats.

In anesthetized, paralyzed and ventilated rats, hypoxia, produced by intratracheal administration of 100% N2 for 20 s, increases sympathetic nerve activity and produces cardiovascular responses. Acute midcollicular decerebration has no effect on these responses in chemo-innervated or chemo-denervated animals. Suprapontine neural structures are, therefore, not required for the rapid sympathetic and cardiovascular responses to acute hypoxia. The results support the view that sympathoexcitatory responses to acute hypoxia depend entirely on the functions of reticulospinal sympathoexcitatory vasomotor neurons of the rostral ventrolateral medulla (RVL).

GABA-mediated inhibition of pacemaker neurons of rostral ventrolateral medulla by clonidine in vitro.

In vitro, clonidine (1, 10, or 30 microM) dose-dependently and reversibly inhibited tonically active pacemaker neurons that correspond to the relatively fast-conducting reticulospinal vasomotor neurons of the rostral ventrolateral reticular nucleus of the medulla oblongata in rats. The clonidine-induced membrane-hyperpolarizing response of these neurons was abolished by either tetrodotoxin, bicuculline, a GABAA receptor antagonist, or 4,4'-diisothiocyano-1,2'-disulphonic stilbene acid, a Cl- channel blocker. We conclude that the clonidine-induced inhibition of the pacemaker neurons of the rostral ventrolateral reticular nucleus is indirect, mediated by synaptic release of gamma-aminobutyric acid (GABA) or GABA-like substances, which activate Cl- channels of the pacemaker neurons of the rostral ventrolateral reticular nucleus.

NMDA receptor-mediated sympathetic chemoreflex excitation of RVL-spinal vasomotor neurones in rats.

1. We investigated the role of N-methyl-D-aspartic acid (NMDA) receptors in mediating hypoxic excitation of the reticulospinal vasomotor neurones of the rostroventrolateral reticular nucleus (RVL) of the medulla oblongata in paralysed ventilated rats. 2. Unilateral close arterial injection of sodium cyanide (100 nmol) into the carotid sinus region or ventilation with 100% N2 for 12 s rapidly, reversibly and reproducibly excited the RVL-spinal vasomotor neurones, followed about 1-2 s later by increases in sympathetic nerve activity and arterial pressure, effects abolished by denervation of the ipsilateral carotid sinus nerve. 3. Ionophoresis onto the RVL-spinal vasomotor neurones of kynurenate (a wide-spectrum antagonist of the excitatory amino acid receptors) or of 2-amino-5-monophosphovaleric acid (APV; a selective NMDA receptor antagonist), but not of xanthurenate (an inactive analogue of kynurenate), blocked the excitation elicited by intracarotid cyanide or 12 s of hypoxia. Kynurenate completely and APV partially blocked the excitatory responses to ionophoretically applied L-glutamate. APV, however, did not alter the excitatory responses of the vasomotor neurones to ionophoreses of kainate and quisqualate. 4. Bilateral microinjection of kynurenate (10 nmol, 50 nl per site) or APV (5 nmol, 50 nl per site) into the RVL blocked the increases in arterial pressure elicited by intracarotid cyanide or 12 s of 100% N2 ventilation. 5. Twenty seconds of intratracheal administration of 100% N2 resulted in complex and prolonged elevations of arterial pressure, the late component of which was affected neither by sinus denervation nor by microinjections of kynurenate or APV into the RVL. 6. We conclude that the sympathetic and cardiovascular responses to stimulation of arterial chemoreceptors result from excitation of RVL-spinal vasomotor neurones via activation of the NMDA subtypes of the excitatory amino acid receptors of the neurones. In contrast, the failure of these antagonists to influence the delayed excitation of the RVL-spinal vasomotor neurons by more prolonged exposure to N2 inhalation further supports the view that these neurones are directly stimulated by hypoxia.